#cell proliferation

Animal Tissue Culture 3

MTT assay measures:

MTT assay developed by Tim Mosmann in 1983 measures cell viability and proliferation based on reduction capacity of living cells. Principle uses yellow tetrazolium salt 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide which enters cells via endocytosis and is reduced by mitochondrial succinate dehydrogenase of complex II and cytosolic NAD(P)H-dependent oxidoreductases cleaving tetrazolium ring with electrons from NADH and NADPH producing insoluble purple formazan crystals accumulating intracellularly in mitochondria and lipid droplets. Metabolically incompetent dead cells fail to r

Ref: Mosmann T J Immunol Methods 1983 MTT viability assay; ATCC MTT Cell Proliferation Assay mitochondrial dehydrogenase reduction protocol.

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Animal Tissue Culture 3

An advantage of EdU assay is:

Predominant advantage of EdU assay over BrdU is preservation of cellular architecture because it avoids harsh DNA denaturation. BrdU detection requires unwinding double helix using strong acid, heat, or enzymatic digestion to expose concealed bromo-deoxyuridine epitope for anti-BrdU antibody of large size 150 kDa that cannot access double-stranded DNA. These treatments hydrolyze fluorescent proteins such as GFP, destroy conformational epitopes of surface antigens CD markers, fragment histology, alter light scatter properties in flow cytometry, and distort cell cycle distributions. EdU click de

Ref: Buck et al Biotechniques 2008 EdU advantage no denaturation preserves antigens; Thermo Fisher Click-iT EdU vs BrdU no DNA denaturation needed.

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Animal Tissue Culture 3

EdU assay differs from BrdU assay because it:

EdU, 5-ethynyl-2'-deoxyuridine, retains Watson-Crick pairing but bears terminal alkyne at 5 position instead of bromine. Detection utilizes bioorthogonal click chemistry specifically copper(I)-catalyzed azide-alkyne cycloaddition first described by Sharpless. In this reaction, alkyne on EdU incorporated into DNA reacts with fluorescent azide forming stable 1,2,3-triazole covalent linkage, catalyzed by CuSO4 reduced to Cu(I) by sodium ascorbate, proceeding rapidly in aqueous buffer at room temperature with exquisite specificity. Because fluorescent azide small molecule penetrates duplex DNA, ha

Ref: Salic A & Mitchison TJ PNAS 2008 EdU click chemistry; Kolb et al Click chemistry concept Nobel Prize 2022 CuAAC azide-alkyne.

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Animal Tissue Culture 3

BrdU is a thymidine analog used to detect:

BrdU, 5-bromo-2'-deoxyuridine, is synthetic halogenated analog of thymidine where methyl group at 5 position replaced by bromine atom similar in size allowing Watson-Crick pairing with adenine during DNA synthesis. During S phase, DNA polymerases incorporate BrdU triphosphate from nucleotide pool into nascent strand in place of thymidine without halting polymerase at low micromolar concentrations. Incorporated BrdU detectable by specific anti-BrdU monoclonal antibodies after DNA denaturation exposes epitope, or by mass spectrometry. Assay quantifies fraction of cells actively replicating genom

Ref: Gratzner HG Science 1982 BrdU thymidine analog DNA replication detection; Alberts MBoC Ch.17 DNA synthesis S phase BrdU incorporation.

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Cancer

Merlin (NF2) suppresses proliferation by

Sequestering growth factor receptors, is consistent with established principles of cell signaling, receptor pharmacology and cellular regulation. Experimental measurements of binding parameters, genetic loss-of-function studies and pharmacological interventions all converge on the same interpretation. Related options address neighboring concepts but do not satisfy the precise criterion stated in the question.